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1392/1/25، جلد ۴۳، شماره ۴، صفحات -
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عنوان فارسی |
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چکیده فارسی مقاله |
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کلیدواژههای فارسی مقاله |
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عنوان انگلیسی |
PRODUCTION OF A HUMAN RECOMBINANT ANTIBODY AGAINST SEROTYPE A CANDIDA ALBICANS |
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چکیده انگلیسی مقاله |
After using 3 different generations of antibodies including human and non-human hyperimmune sera, monoclonal antibodies and chimeric antibodies, more recently a newer approach has been developed in which the antibody genes are cloned directly from a patient peripheral B-lymphocytes and expressed in a host like E. coli. In this study the Candida albicans serotype A (NCTC 3153) mannan was purified using a modified Fehling method and used for selection of human recombinant antibody from a C. albicans phage antibody library. After four rounds of affinity selecting (panning), 2 predominant clones were chosen by DNA fingerprinting and ELISA. A 248 amino acid DNA fragment coding for anti-C. albicans mannan scFv was sequenced and cloned in a pBAD-TOPO cloning vector to produce a soluble and phage free antibody. The analysis of antibody sequences by V base Index (DNAPLOT) confirmed the human antibody origin with the VH4 family in V segment of heavy variable chain and VL3 (Lambda 3) in J segment of the light variable chain. This antibody fragment was purified using immobilized metal affinity chromatography and inmmunoblotted as a 31kDa recombinant protein. |
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کلیدواژههای انگلیسی مقاله |
human recombinant antibody, scFv, protein purification, |
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نویسندگان مقاله |
34593--- |
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نشانی اینترنتی |
http://acta.tums.ac.ir/index.php/acta/article/viewArticle/2857 |
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زبان مقاله منتشر شده |
en |
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نوع مقاله منتشر شده |
Article(s) |
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